Effect of stirring speed on the production of phenolic secondary metabolites and growth of Buddleja cordata cells cultured in mechanically agitated bioreactor

  • Alicia Monserrat Vazquez-Marquez
  • Carmen Zepeda-Gómez
  • Cristina Burrola-Aguilar
  • Antonio Bernabé-Antonio
  • Aurelio Nieto-Trujillo
  • Francisco Cruz-Sosa
  • Mario Rodríguez-Monroy
  • María Elena Estrada-ZúñigaEmail author
Original Article


Shake-flask in vitro culture of Buddleja cordata cells produces large amounts of biomass and synthetizes verbascoside (VB), linarin and hydroxycinnamic acids, bioactive phenolic secondary metabolites (PSMs). In this work, we determined the effect of stirring speed on the growth of and production of PSMs [total phenolic, phenylethanoid glycoside and flavonoid contents (PeC, PeGC and FC, respectively)] by B. cordata cells cultured in two bioreactors. Two different stirring speeds (120 and 400 rpm) were tested in two stirred-tank bioreactors: a 2 L bioreactor equipped with a ring diffuser (B2RD) and a 3 L bioreactor with a sintered diffuser (B3SD). Growth kinetics of B. cordata cells were measured in the bioreactors and shake-flask systems. The stirring speed and type of bioreactor affected phases, parameters of growth and production of PSMs. The highest production of biomass (13.62 g L−1) and PSMs [PeC of 64.63 mg gallic acid equivalents g−1 (mg GAE g−1); PeGC of 119.24 mg VB equivalents g−1 (mg VBE g−1); and FC of 5.02 mg quercetin equivalents g−1 (mg QE g−1)] occurred in B2RD at 400 rpm. These values were similar to the found in shake-flasks system. This work establishes the basis for bioprocess advances of B. cordata focused on the development of a sustainable strategy for the management of natural resources and as a source of bioactive PSMs on a large scale.

Key message

Buddleja cordata cells cultured in a mechanically agitated bioreactor possess an outstanding biosynthetic potential that represents a suitable biotechnological alternative for the production of bioactive phenolic secondary metabolites.


Bioreactor Buddleja cordata Phenolic secondary metabolites Diffuser Stirring speed Verbascoside 



Specific growth rate


Analysis of variance


2 L Bioreactor equipped with a ring diffuser


3 L Bioreactor equipped with a sintered diffuser


Cell suspension culture of B. cordata


Cellular viability


Dissolved oxygen


Dry weight


Flavonoid (s)


Total flavonoid content


Growth index


Gallic acid equivalent


Maximum biomass


Murashige and Skoog-modified culture medium


Phenolic compound (s)


Total phenolic content


Phenylethanoid glycoside (s)


Total phenylethanoid glycoside content


Phenolic secondary metabolite (s)


Quercetin equivalent


Specific production rate of total flavonoids


Specific production rate of total phenolics


Specific production rate of total phenylethanoid glycosides


Rosmarinic acid


Secondary metabolite (s)


Doubling time


Total sugar content




Verbascoside equivalent


Yield of biomass from substrate



The authors thank the Universidad Autónoma del Estado de México (UAEM) for financing this thesis project through the Programa de Investigación Científica, Innovación y Desarrollo UAEM 2014 (Project No. 3742/2014/CIB: Desarrollando Avances Biotecnológicos Sobre la Producción de Verbascósido por Cultivos de Células de Buddleja cordata) and to the Consejo Nacional de Ciencia y Tecnología (CONACyT) through grant number 620491 for the Master’s studies of AMV-M at the Posgrado en Ciencias Agropecuarias y Recursos Naturales from UAEM.

Authors contributions

AMV-M, as a Master student and for her thesis project, participated in all the experimental work, analysis and interpretation of data, and writing of this manuscript. CZ-G supervised the establishment of the experiments and provided chemical standards. CB-A participated in the standardization of analytical procedures to quantify the secondary metabolites. AB-A participated in the analysis of the growth kinetics and their statistical analysis. AN-T participated in the experimental work on phytochemical analysis. FC-S contributed to the design and execution of experiments on shake-flask cultures; participated in the analysis and interpretation of data. MR-M contributed to the thesis project of MAV-M as an assessor; supervised the establishment of bioreactor in vitro cultures; contributed to the planning and execution of this project, which led to this publication; and contributed to the preparation and writing of this manuscript. MEE-Z contributed to the design, execution and direction of all the experiments of this project, as she was the thesis project director of AMV-M; was the responsible for the design and direction of Project No. 3742/2014/CIB, which financially supported this thesis project; and critically contributed to the preparation of the manuscript until approving the final submitted version. All authors critically reviewed the manuscript and approved the final version.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Facultad de CienciasUniversidad Autónoma del Estado de MéxicoTolucaMexico
  2. 2.Centro de Investigación en Recursos Bióticos-Facultad de CienciasUniversidad Autónoma del Estado de MéxicoTolucaMexico
  3. 3.Departamento de Madera, Celulosa y Papel, Centro Universitario de Ciencias Exactas e IngenieríasUniversidad de GuadalajaraZapopanMexico
  4. 4.Departamento de BiotecnologíaUniversidad Autónoma Metropolitana-IztapalapaCol. VicentinaMexico
  5. 5.Departamento de BiotecnologíaCentro de Desarrollo de Productos Bióticos del Instituto Politécnico Nacional, YautepecMorelosMexico

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